The adaptor protein Ruk/CIN85 activates plasminogen activator inhibitor-1 (PAI-1) expression via hypoxia-inducible factor-1α

 

 Buy Article Permissions and Reprints

Summary

Increased levels of plasminogen activator inhibitor-1 (PAI-1) indicate an enhanced risk of ischaemic/hypoxic cardiovascular events and a poor prognosis. The expression of PAI-1 can be induced by various stimuli including hypoxia, insulin and insulin-like growth factor 1 (IGF-1). The hypoxia-inducible factor-1 (HIF-1) is critical for hypoxia or insulin/IGF-1 mediated PAI-1 induction, but the components involved in merging the signals are not known so far. The adaptor/scaffold protein Ruk/CIN85 may be a candidate since it plays important roles in the regulation of processes associated with cardiovascular and oncological diseases such as downregulation of receptor tyrosine kinases, apoptosis, adhesion and invasion. Therefore, it was the aim of this study to investigate the involvement of Ruk/CIN85 in the regulation of PAI-1 expression. It was found that Ruk/CIN85 induced PAI-1 mRNA and protein expression both under normoxia and hypoxia. The induction of PAI-1 expression by Ruk/CIN85 occurred at the transcriptional level since the half-life of PAI-1 mRNA was not affected in cells overexpressing Ruk/ CIN85 and reporter gene assays using wild-type and mutant human PAI-1 promoter luciferase constructs showed that the hypoxia responsive element was responsible for Ruk/CIN85 effects. Further, knocking down HIF-1α abolished not only the hypoxia-dependent but also the Ruk/CIN85-dependent PAI-1 induction. In addition, transient or stable overexpression of Ruk/CIN85 also induced HIF-1α protein levels and HIF-1 activity and knocking down Ruk/CIN85 reversed these effects. Thereby, Ruk/CIN85 interfered with the proline hydroxylation-dependent HIF-1α protein destabilisation. Together, these results provide the first evidence that Ruk/CIN85 induces PAI-1 expression via modulation of HIF-1α stability.

• References

• 1 Kruithof EK, Tran TC, Bachmann F. The fast-acting inhibitor of tissue-type plasminogen activator in plasma is also the primary plasma inhibitor of urokinase. Thromb Haemost 1986; 55: 65-69.
  Article in Thieme ConnectPubMedGoogle Scholar
• 2 Erickson LA, Hekman CM, Loskutoff DJ. The primary plasminogen-activator inhibitors in endothelial cells, platelets, serum, and plasma are immunologically related. Proc Natl Acad Sci USA 1985; 82: 8710-8714.
  CrossrefPubMedGoogle Scholar
• 3 Reilly CF, McFall RC. Platelet-derived growth factor and transforming growth factor-beta regulate plasminogen activator inhibitor-1 synthesis in vascular smooth muscle cells. J Biol Chem 1991; 266: 9419-9427.
  PubMedGoogle Scholar
• 4 Heaton JH, Nebes VL, O’Dell LG. et al. Glucocorticoid and cyclic nucleotide regulation of plasminogen activator and plasminogen activator-inhibitor gene expression in primary cultures of rat hepatocytes. Mol Endocrinol 1989; 03: 185-192.
  CrossrefPubMedGoogle Scholar
• 5 Busso N, Nicodeme E, Chesne C. et al. Urokinase and type I plasminogen activator inhibitor production by normal human hepatocytes: modulation by inflammatory agents. Hepatology 1994; 20: 186-190.
  PubMedGoogle Scholar
• 6 Eren M, Painter CA, Atkinson JB. et al. Age-dependent spontaneous coronary arterial thrombosis in transgenic mice that express a stable form of human plasminogen activator inhibitor-1. Circulation 2002; 106: 491-496.
  CrossrefPubMedGoogle Scholar
• 7 De Taeye B, Smith LH, Vaughan DE. Plasminogen activator inhibitor-1: a common denominator in obesity, diabetes and cardiovascular disease. Curr Opin Pharmacol 2005; 05: 149-154.
  CrossrefPubMedGoogle Scholar
• 8 Decock J, Paridaens R, Cufer T. Proteases and metastasis: clinical relevance nowadays?. Curr Opin Oncol 2005; 17: 545-550.
  CrossrefPubMedGoogle Scholar
• 9 Ha H, Oh EY, Lee HB. The role of plasminogen activator inhibitor 1 in renal and cardiovascular diseases. Nat Rev Nephrol 2009; 05: 203-211.
  CrossrefPubMedGoogle Scholar
• 10 Schneider DJ, Sobel BE. Augmentation of synthesis of plasminogen activator inhibitor type 1 by insulin and insulin-like growth factor type I: implications for vascular disease in hyperinsulinemic states. Proc Natl Acad Sci USA 1991; 88: 9959-9963.
  CrossrefPubMedGoogle Scholar
• 11 Dimova EY, Kietzmann T. Metabolic, hormonal and environmental regulation of plasminogen activator inhibitor-1 (PAI-1) expression: lessons from the liver. Thromb Haemost 2008; 100: 992-1006.
  Article in Thieme ConnectPubMedGoogle Scholar
• 12 Kietzmann T, Roth U, Jungermann K. Induction of the plasminogen activator inhibitor-1 gene expression by mild hypoxia via a hypoxia response element binding the hypoxia-inducible factor-1 in rat hepatocytes. Blood 1999; 94: 4177-4185.
  PubMedGoogle Scholar
• 13 Kietzmann T, Samoylenko A, Roth U. et al. Hypoxia-inducible factor-1 and hypoxia response elements mediate the induction of plasminogen activator inhibitor-1 gene expression by insulin in primary rat hepatocytes. Blood 2003; 101: 907-914.
  CrossrefPubMedGoogle Scholar
• 14 Dimova EY, Moller U, Herzig S. et al. Transcriptional regulation of plasminogen activator inhibitor-1 expression by insulin-like growth factor-1 via MAP kinases and hypoxia-inducible factor-1 in HepG2 cells. Thromb Haemost 2005; 93: 1176-1184.
  Article in Thieme ConnectPubMedGoogle Scholar
• 15 Dimova EY, Kietzmann T. The MAPK pathway and HIF-1 are involved in the induction of the human PAI-1 gene expression by insulin in the human hepatoma cell line HepG2. Ann N Y Acad Sci 2006; 1090: 355-367.
  CrossrefPubMedGoogle Scholar
• 16 Flynn DC. Adaptor proteins. Oncogene 2001; 20: 6270-6272.
  CrossrefPubMedGoogle Scholar
• 17 Gout I, Middleton G, Adu J. et al. Negative regulation of PI 3-kinase by Ruk, a novel adaptor protein. EMBO J 2000; 19: 4015-4025.
  CrossrefPubMedGoogle Scholar
• 18 Szymkiewicz I, Kowanetz K, Soubeyran P. et al. CIN85 participates in Cbl-b-mediated down-regulation of receptor tyrosine kinases. J Biol Chem 2002; 277: 39666-39672.
  CrossrefPubMedGoogle Scholar
• 19 Havrylov S, Ichioka F, Powell K. et al. Adaptor protein Ruk/CIN85 is associated with a subset of COPI-coated membranes of the Golgi complex. Traffic 2008; 09: 798-812.
  CrossrefPubMedGoogle Scholar
• 20 Hutchings NJ, Clarkson N, Chalkley R. et al. Linking the T cell surface protein CD2 to the actin-capping protein CAPZ via CMS and CIN85. J Biol Chem 2003; 278: 22396-22403.
  CrossrefPubMedGoogle Scholar
• 21 Raab RM, Bullen J, Kelleher J. et al. Regulation of mouse hepatic genes in response to diet induced obesity, insulin resistance and fasting induced weight reduction. Nutr Metab (Lond) 2005; 02: 15.
  CrossrefPubMedGoogle Scholar
• 22 Chen B, Borinstein SC, Gillis J. et al. The glioma-associated protein SETA interacts with AIP1/Alix and ALG-2 and modulates apoptosis in astrocytes. J Biol Chem 2000; 275: 19275-19281.
  CrossrefPubMedGoogle Scholar
• 23 Liu Q, Berchner-Pfannschmidt U, Moller U. et al. A Fenton reaction at the endoplasmic reticulum is involved in the redox control of hypoxia-inducible gene expression. Proc Natl Acad Sci USA 2004; 101: 4302-4307.
  CrossrefPubMedGoogle Scholar
• 24 Immenschuh S, Hinke V, Ohlmann A. et al. Transcriptional activation of the haem oxygenase-1 gene by cGMP via a cAMP response element/activator protein-1 element in primary cultures of rat hepatocytes. Biochem J 1998; 334: 141-146.
  CrossrefPubMedGoogle Scholar
• 25 Bonello S, Zahringer C, BelAiba RS. et al. Reactive oxygen species activate the HIF-1alpha promoter via a functional NFkappaB site. Arterioscler Thromb Vasc Biol 2007; 27: 755-761.
  CrossrefPubMedGoogle Scholar
• 26 Ganjam GK, Dimova EY, Unterman TG. et al. FoxO1 and HNF-4 are involved in regulation of hepatic glucokinase gene expression by resveratrol. J Biol Chem 2009; 284: 30783-30797.
  CrossrefPubMedGoogle Scholar
• 27 Mayevska O, Shuvayeva H, Igumentseva N. et al. Expression of adaptor protein Ruk/CIN85 isoforms in cell lines of various tissue origins and human melanoma. Exp Oncol 2006; 28: 275-281.
  PubMedGoogle Scholar
• 28 Fattal PG, Schneider DJ, Sobel BE. et al. Post-transcriptional regulation of expression of plasminogen activator inhibitor type 1 mRNA by insulin and insulin-like growth factor 1. J Biol Chem 1992; 267: 12412-12415.
  PubMedGoogle Scholar
• 29 Uchiyama T, Kurabayashi M, Ohyama Y. et al. Hypoxia induces transcription of the plasminogen activator inhibitor-1 gene through genistein-sensitive tyrosine kinase pathways in vascular endothelial cells. Arterioscler Thromb Vasc Biol 2000; 20: 1155-1161.
  CrossrefPubMedGoogle Scholar
• 30 Tanimoto K, Makino Y, Pereira T. et al. Mechanism of regulation of the hypoxiainducible factor-1 alpha by the von Hippel-Lindau tumor suppressor protein. EMBO J 2000; 19: 4298-4309.
  CrossrefPubMedGoogle Scholar
• 31 Pugh CW, O’Rourke JF, Nagao M. et al. Activation of hypoxia-inducible factor-1; definition of regulatory domains within the alpha subunit. J Biol Chem 1997; 272: 11205-11214.
  CrossrefPubMedGoogle Scholar
• 32 Ivan M, Kondo K, Yang H. et al. HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. Science 2001; 292: 464-468.
  CrossrefPubMedGoogle Scholar
• 33 Huang J, Zhao Q, Mooney SM. et al. Sequence determinants in hypoxia-inducible factor-1alpha for hydroxylation by the prolyl hydroxylases PHD1, PHD2, and PHD3. J Biol Chem 2002; 277: 39792-39800.
  CrossrefPubMedGoogle Scholar
• 34 Pereira T, Zheng X, Ruas JL. et al. Identification of residues critical for regulation of protein stability and the transactivation function of the hypoxia-inducible factor-1a by the von Hippel-Lindau tumor suppressor gene product. J Biol Chem 2002; 278: 6816-6823.
  PubMedGoogle Scholar
• 35 Lando D, Peet DJ, Whelan DA. et al. Asparagine hydroxylation of the HIF transactivation domain a hypoxic switch. Science 2002; 295: 858-861.
  CrossrefPubMedGoogle Scholar
• 36 Jiang BH, Zheng JZ, Leung SW. et al. Transactivation and inhibitory domains of hypoxia-inducible factor 1alpha. Modulation of transcriptional activity by oxygen tension. J Biol Chem 1997; 272: 19253-19260.
  CrossrefPubMedGoogle Scholar
• 37 Ema M, Hirota K, Mimura J. et al. Molecular mechanisms of transcription activation by HLF and HIF1alpha in response to hypoxia: their stabilization and redox signal-induced interaction with CBP/p300. EMBO J 1999; 18: 1905-1914.
  CrossrefPubMedGoogle Scholar
• 38 Dellas C, Loskutoff DJ. Historical analysis of PAI-1 from its discovery to its potential role in cell motility and disease. Thromb Haemost 2005; 93: 631-640.
  Article in Thieme ConnectPubMedGoogle Scholar
• 39 Pinsky DJ, Liao H, Lawson CA. et al. Coordinated induction of plasminogen activator inhibitor-1 (PAI-1) and inhibition of plasminogen activator gene expression by hypoxia promotes pulmonary vascular fibrin deposition. J Clin Invest 1998; 102: 919-928.
  CrossrefPubMedGoogle Scholar
• 40 Fink T, Kazlauskas A, Poellinger L. et al. Identification of a tightly regulated hypoxia-response element in the promoter of human plasminogen activator inhibitor-1. Blood 2002; 99: 2077-2083.
  CrossrefPubMedGoogle Scholar
• 41 Bruick RK, McKnight SL. A conserved family of prolyl-4-hydroxylases that modify HIF. Science 2001; 294: 1337-1340.
  CrossrefPubMedGoogle Scholar
• 42 Tibaldi EV, Reinherz EL. CD2BP3, CIN85 and the structurally related adaptor protein CMS bind to the same CD2 cytoplasmic segment, but elicit divergent functional activities. Int Immunol 2003; 15: 313-329.
  CrossrefPubMedGoogle Scholar
• 43 Havrylov S, Rzhepetskyy Y, Malinowska A. et al. Proteins recruited by SH3 domains of Ruk/CIN85 adaptor identified by LC-MS/MS. Proteome Sci 2009; 07: 21.
  CrossrefPubMedGoogle Scholar
• 44 Buchman VL, Luke C, Borthwick EB. et al. Organization of the mouse Ruk locus and expression of isoforms in mouse tissues. Gene 2002; 295: 13-17.
  CrossrefPubMedGoogle Scholar
• 45 Dikic I. CIN85/CMS family of adaptor molecules. FEBS Lett 2002; 529: 110-115.
  CrossrefPubMedGoogle Scholar
• 46 Hopfer U, Hopfer H, Jablonski K. et al. The novel WD-repeat protein Morg1 acts as a molecular scaffold for hypoxia-inducible factor prolyl hydroxylase 3 (PHD3). J Biol Chem 2006; 281: 8645-8655.
  CrossrefPubMedGoogle Scholar
• 47 Ravichandran KS. Signaling via Shc family adapter proteins. Oncogene 2001; 20: 6322-6330.
  CrossrefPubMedGoogle Scholar
• 48 Jung F, Haendeler J, Hoffmann J. et al. Hypoxic induction of the hypoxia-inducible factor is mediated via the adaptor protein Shc in endothelial cells. Circ Res 2002; 91: 38-45.
  CrossrefPubMedGoogle Scholar
• 49 Harbeck N, Kates RE, Gauger K. et al. Urokinase-type plasminogen activator (uPA) and its inhibitor PAI-I: novel tumor-derived factors with a high prognostic and predictive impact in breast cancer. Thromb Haemost 2004; 91: 450-456.
  Article in Thieme ConnectPubMedGoogle Scholar
• 50 Semenza GL. Regulation of cancer cell metabolism by hypoxia-inducible factor 1. Semin Cancer Biol 2009; 19: 12-16.
  CrossrefPubMedGoogle Scholar